How do you determine the size of each protein with this data? length of column x πr2 or riboflavin elution , Elution volume is related to size! Standard lnes (again) , Quick estimate: Log(Mr) vs Ve/V。 Better: Log(M) vs (Ve-V/(VVo) You should use the better method Your results Protein eluted 09 0.7 0.6 0.5 4 Standards: 110 kDa V.-10 mL 65 kDa V 18 mL 45 kDa = 25 mL 32 kDa V 33 mL 0.3 Dextran V o Riboflavin V 51 mL 13 579 1113 5 17 19 23 23 25 27 29 33 33 35 37 39 41 43 45 67 495 Fraction Number amL fractions cellected length of column x πr2 or riboflavin elution , Elution volume is related to size! Standard lnes (again) , Quick estimate: Log(Mr) vs Ve/V。 Better: Log(M) vs (Ve-V/(VVo) You should use the better method Your results Protein eluted 09 0.7 0.6 0.5 4 Standards: 110 kDa V.-10 mL 65 kDa V 18 mL 45 kDa = 25 mL 32 kDa V 33 mL 0.3 Dextran V o Riboflavin V 51 mL 13 579 1113 5 17 19 23 23 25 27 29 33 33 35 37 39 41 43 45 67 495 Fraction Number amL fractions cellected
This is a typical profile of the gel purification process. First, the highermolecular weight protein elutes. The lower the elution volume, the higher the weight of the molecule. A standard graph of proteins with known weight and elution volume is plotted onto which the unknown one can be mapped to find its relative molecular mass.